Pfizer Worldwide Research and Development , Eastern Point Road , Groton , Connecticut 06340 , United States.
Pfizer Worldwide Research and Development , 10770 Science Center Drive , La Jolla , California 92121 , United States.
J Med Chem. 2018 Apr 26;61(8):3626-3640. doi: 10.1021/acs.jmedchem.8b00116. Epub 2018 Apr 9.
In this report, we describe a method whereby lead molecules can be converted into several new analogues each using liver microsomes. Less than one micromole of substrate is incubated with liver microsomes (mouse, rat, hamster, guinea pig, rabbit, dog, monkey, or human) to produce multiple products which are isolated and analyzed by quantitative cryomicroprobe NMR (qNMR) spectroscopy. The solutions from qNMR analysis were then used as stocks that were diluted into biochemical assays. Nine human phosphodiesterase-2 (PDE2) inhibitors yielded 36 new analogues. Products were tested for PDE2 inhibition, intrinsic clearance in human hepatocytes, and membrane permeability. Two of the products (2c and 4b) were 3-10× more potent than their respective parent compounds and also had improved metabolic stability. Others offered insights into structure-activity relationships. Overall, this process of using liver microsomes at a submicromole scale of substrate is a useful approach to rapid and cost-effective late-stage lead diversification.
在本报告中，我们描述了一种方法，通过该方法可以使用肝微粒体将先导化合物转化为几种新的类似物。用不到 1 微摩尔的底物与肝微粒体（鼠、大鼠、仓鼠、豚鼠、兔、狗、猴或人）孵育以产生多种产物，然后通过定量 cryomicroprobe NMR（qNMR）光谱法对其进行分离和分析。qNMR 分析的溶液随后被用作储备液，然后将其稀释到生化测定中。九种人磷酸二酯酶-2（PDE2）抑制剂产生了 36 种新类似物。对产物进行了 PDE2 抑制、人肝细胞内固有清除率和膜通透性测试。其中两种产物（2c 和 4b）比其各自的母体化合物的效力高 3-10 倍，并且代谢稳定性也得到了改善。其他产物则提供了对构效关系的深入了解。总的来说，这种在亚微摩尔底物规模下使用肝微粒体的方法是一种快速且具有成本效益的后期先导化合物多样化的有用方法。
